Entacapone for prevention and treatment of obesity and related metabolic diseases

ABSTRACT

Obesity is inhibited by administering to a person in need thereof an effective amount of entacapone ((2E)-2-cyano-3-(3,4-dihydroxy-5-nitrophenyl)-N,N-diethylprop-2-enamide), or a pharmaceutically-acceptable salt thereof, particularly in conjunction with a second, different anti-obesity medicament. Pharmaceutical compositions comprise entacapone copackaged or coformulated with a second, different anti-obesity medicament.

INTRODUCTION

Obesity is becoming a severe health problem worldwide and many factorscontribute to this chronic disease, including environmental factors andgenetic factors. Recently, GWAS (genome-wide association studies) wereapplied to investigate patients with obesity and a single gene—FTO (fatmass and obesity) was identified to strongly associate with obesity.¹⁻⁴FTO's functional role in obesity was subsequently confirmed intransgenetic animal models, such as FTO knockout mouse,FTO-overexpression mouse and FTO-I367F mutation mouse.⁵⁻⁸ Morespecifically, FTO global-knockout and neuron-specific knockout inducebody weight loss^(5,6), while FTO gene overexpression results inobesity.⁸ One mis-sense mutation was observed to inhibit FTO enzymaticfunction and protect mouse from obesity.⁷ Nevertheless, FTO is expressedin many tissues, especially in hypothalamic nuclei controlling energyexpenditure,⁹ which is consistent with the suggestion that FTO affectsenergy homeostasis.⁶

FTO protein is an α-ketoglutarate and iron (II) dependent nucleic aciddemethylase.^(9,10) Its preferred substrate is N⁶-meA in message RNA,which locates near the stop codon and influences genetranslation.^(11,12) However, its mode of mechanism is not clear, suchas its upstream and downstream effectors and its regulation mechanism.It is desirable to identify small molecule, drug-like FTO inhibitors forstudying FTO function and discovering novel anti-obesity agent.

Existing drugs are approved for human use and have known safetyprofiles, so they can be rapidly evaluated for new indications ofbiological interest. Herein, we report the identification of a known FDAapproved drug—Entacapone as FTO inhibitor using structure-based virtualscreening method in combination with biological activity measurements,including enzymatic activity, cellular activity and in high-fat dietinduced obesity (DIO) animal model.

SUMMARY OF THE INVENTION

The invention provides methods and composition for inhibiting weightgain, wherein the methods are also applicable to promoting weight loss,reducing serum LDL, cholesterol, LDL-c, and/or triglycerides, and/ortreating an obesity related or metabolic disease or ameliorating orreducing the pathology or severity of an obesity related or metabolicdisease or a symptom of an obesity related or metabolic disease selectedfrom diabetes, hyperglycemia, diabetic nephropathy, hyperlipemia,coronary heart disease, atherosclerosis, hypertension, cardiovascular orcerebrovascular diseases, or liver, kidney or thyroid diseases.

In one aspect the invention provides methods of inhibiting weight gain,promoting weight loss, reducing serum LDL, cholesterol, LDL-c, ortriglycerides, or treating atherosclerosis, comprising administering toa person in need thereof an effective amount of entacapone((2E)-2-cyano-3-(3,4-dihydroxy-5-nitrophenyl)-N,N-diethylprop-2-enamide),or a pharmaceutically-acceptable salt thereof.

In another aspect the invention provides methods of inhibiting weightgain comprising administering to a person in need thereof an effectiveamount of entacapone in conjunction with one or more differentmedicaments for inhibiting weight gain, promoting weight loss, reducingserum LDL, cholesterol, LDL-c, or triglycerides, or treatingatherosclerosis.

In particular embodiments thereof, the person meets one or morecriteria, such as (a) is not diagnosed with Parkinson's disease; (b) isless than 50, 40 or 30 years old; (c) is obese or over-weight; (d)suffers from or is diagnosed with an obesity related disease selectedfrom diabetes, hyperglycemia, diabetic nephropathy, hyperlipemia,coronary heart disease, atherosclerosis, hypertension, cardiovascular orcerebrovascular diseases, or liver, kidney or thyroid diseases; (e) hasgenotype: SNP rs7202116 (G), rs1421085 (C), or rs9939609 (A); and/or (f)pathogenically expresses or over-expresses FTO or Fto.

In particular embodiments the method further comprising detecting in theperson one or more of the criteria, particularly genotype SNP rs7202116(G), or over-expression of FTO or Fto.

In particular embodiments the method further comprises detecting aresultant inhibition of weight gain, promotion of weight loss, and/orimprovement or amelioration of one or more of the criteria.

In particular embodiments, the method further comprises administering tothe person an effective amount of one or more additional, differentmedicaments for inhibiting weight gain, promoting weight loss, reducingserum LDL, cholesterol, LDL-c, or triglycerides, or treatingatherosclerosis, particularly wherein the entacapone and medicament(s)are copackaged, coformulated or coadministered.

In another aspect the invention provides pharmaceutical compositionscomprising entacapone copackaged or coformulated with one or more of theadditional, different medicaments for inhibiting weight gain, promotingweight loss, reducing serum LDL, cholesterol, LDL-c, or triglycerides,or treating atherosclerosis.

In particular embodiments the subject methods and compositions employentacapone dosages of 0.25-5 g, or g/day, or 0.5-5 g, or g/day.

The invention encompasses all combination of the particular embodimentsrecited herein.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. The chemical structure of entacapone.

FIG. 2. The dose-dependent inhibition of entacapone against FTO proteinmeasured in demethylation assay.

FIG. 3. The dose-dependent inhibition activity of entacapone ontriglyceride synthesis in Huh-7 cell line. The X-axis is the compoundconcentration. The Y-axis is “read-out” of the triglycerideconcentration using dyeing method.

FIG. 4. Effects of entacapone on body weight in rats. (*p-value<0.05)

FIG. 5. Effects of entacapone on food intake in rats.

FIG. 6. Effects of entacapone on LDL-c of serum in rats.(***p-value<0.005)

FIG. 7. Effects of entacapone on adipose and hepatic tissues in rats.

FIG. 8. The anti-atherosclerosis efficacy of entacapone inLdlr-Deficient Mice. a and b. The Oil red O-stained lesion area inaortic sinuses of control group and drug-treated group, respectively. c.Quantitative analysis of lesional areas in the aortic sinuses of twogroups.

FIG. 9. Effects of entacapone on body weight in obese mice.(*p-value<0.05)

DESCRIPTION OF PARTICULAR EMBODIMENTS OF THE INVENTION

In one aspect the invention provides methods of inhibiting weight gainor promoting weight loss comprising administering to a person in needthereof an effective amount of entacapone((2E)-2-cyano-3-(3,4-dihydroxy-5-nitrophenyl)-N,N-diethylprop-2-enamide),or a pharmaceutically-acceptable salt thereof, and includingstereoisomers, particularly E-Z isomers, including polymeric forms,particularly the A form of the E isomer (e.g. U.S. Pat. No. 5,135,950).

The subject methods and compositions are also applicable to relatedindications: promoting weight loss, reducing serum LDL, cholesterol,LDL-c, and/or triglycerides, and/or treating an obesity related diseaseor ameliorating or reducing the pathology or severity of an obesityrelated disease or a symptom of an obesity related disease selected fromdiabetes, hyperglycemia, diabetic nephropathy, hyperlipemia, coronaryheart disease, atherosclerosis, hypertension, cardiovascular orcerebrovascular diseases, or liver, kidney or thyroid diseases.Accordingly, the invention provides methods and compositions whereinentacapone is copackaged, coformulated or coadministered with one ormore different medicaments for, specific for, or indicated for theserelated indications, and the methods may further comprising detecting,determining or diagnosing in the person one or more of the indications,and/or detecting a resultant improvement or amelioration ofcorresponding condition or symptom in the person.

In one aspect the invention provides methods of inhibiting weight gaincomprising administering to a person in need thereof an effective amountof entacapone in conjunction with one or more different medicaments forinhibiting weight gain, promoting weight loss, reducing serum LDL,cholesterol, LDL-c, or triglycerides, or treating atherosclerosis,particularly wherein the entacapone and medicament(s) are copackaged,coformulated or coadministered.

Preferred different medicaments for these indications include differentanti-weight gain medicaments, particularly a food intake inhibitorand/or a food absorption inhibitor; particularly Orlistat, Sibutramine,Lorcaserin, Rimonabant, Metformin, Exenatide, Pramlintide, or apharmaceutically-acceptable salt thereof, and cholesterol loweringdrugs, such as statins, including atorvastatin (Lipitor), fluvastatin(Lescol), lovastatin (Altoprev, Mevacor), pravastatin (Pravachol),rosuvastatin (Crestor), simvastatin (Zocor), or apharmaceutically-acceptable salt thereof.

In particular embodiments thereof, the person meets one or more criteriaindicative of the disclosed non-Parkinson's indication, and the methodmay further comprising detecting in the person one or more of thecriteria, and/or detecting a resultant inhibition of weight gain,promotion of weight loss and/or improvement or amelioration of one ormore of such criteria.

Criteria indicative of the disclosed non-Parkinson's indication includewherein the person (a) is not suffering from, or is not diagnosed withParkinson's disease or symptoms thereof or other prior indication forentacapone or is not suffering from, or is not diagnosed with anydegenerative disease of the central nervous system; (b) is less than 50,40 or 30 years old; (c) is over-weight (e.g. a BMI of 25-30) or obese(e.g. a BMI of over 30); (d) suffers from, or is diagnosed with anobesity related disease selected from diabetes, hyperglycemia, diabeticnephropathy, hyperlipemia, coronary heart disease, atherosclerosis,hypertension, cardiovascular or cerebrovascular diseases, or liver,kidney or thyroid diseases; or (e) has a genotype associated withobesity or pathogenic or medically-undesirable weight gain, such as SNPrs7202116 (G), rs1421085 (C), or rs9939609 (A), or a surrogate or proxySNP in linkage disequilibrium therewith (with respect to the correlativephenotype; see references below) and having a r² value greater than 0.5;and/or (f) pathogenically expresses or over-expresses FTO or Fto (e.g.comprises and expresses a multi-copy fto gene). Re rs7202116 G, see e.g.Yang et al., FTO genotype is associated with phenotypic variability ofbody mass index, Nature, Sep. 16, 2012, doi: 10.1038/nature11401 [epub];re rs9939609 A, see e.g. Freathy R M, et al (2008). “Common variation inthe FTO gene alters diabetes-related metabolic traits to the extentexpected, given its effect on BMI”. Diabetes 57 (5): 1419-26.doi:10.2337/db07-1466. PMC 3073395. PMID 18346983; re rs1421085 C, seee.g. Dina C, et al., (2007). “Variation in FTO contributes to childhoodobesity and severe adult obesity”. Nature Genetics 39 (6): 724-6.doi:10.1038/ng2048. PMID 17496; and for multi-copy fto gene mouse, seee.g. Church et al., Overexpression of Fto leads to increased food intakeand results in obesity, Nature Genetics, published online 14 Nov. 2010,doi:10.1038/ng.713.

In particular embodiments the person does not suffer from a pathogenicdeficiency of L-DOPA (L-3,4-dihydroxyphenylalanine), does not indicateL-DOPA, and/or is not in need of L-DOPA (levopoda) or a dopaminergicagent, and/or the entacapone is not administered in conjunction withL-DOPA or a doaminergic agent.

In particular embodiments, the method further comprises administering tothe person an effective amount of one or more additional, differentmedicament for inhibiting weight gain, promoting weight loss, reducingserum LDL, cholesterol, LDL-c, or triglycerides, or treatingatherosclerosis, particularly wherein the entacapone and medicament(s)are copackaged, coformulated or coadministered.

In another aspect the invention provides pharmaceutical compositionscomprising entacapone copackaged or coformulated with one or more of theadditional, different medicaments for inhibiting weight gain, promotingweight loss, reducing serum LDL, cholesterol, LDL-c, or triglycerides,or treating atherosclerosis, particularly different anti-weight gainmedicaments. For example, the entacapone and additional, differentanti-weight gain medicaments may be coformulated, particularly in unitdosage form, or unit dosage forms of each may be copackaged in amultipack adapted for sequential use, such as blisterpack, comprisingsheets of unit dosage forms. Exemplary coformulation and copackagingsare shown in Table 2.

Table 2. Exemplary Coformulation and Copackagings.

1. Entacapone/Orlistat in 1000 mg/120 mg or 2000 mg/120 mg coformulatedtablets.2. Entacapone/Sibutramine in 1000 mg/10 mg or 2000 mg/10 mg coformulatedtablets.3. Entacapone/Lorcaserin in 1000 mg/10 mg or 2000 mg/10 mg coformulatedtablets.4. Entacapone/Rimonabant in 1000 mg/20 mg or 2000 mg/20 mg coformulatedtablets.5. Entacapone/Metformin in 1000 mg/500 mg or 2000 mg/500 mg coformulatedtablets.6. Entacapone/Exenatide: 1000 mg or 2000 mg tablet/250 mcg/mL solution(Byetta), copackaged7. Entacapone/Exenatide: 1000 mg or 2000 mg tablet/suspension powder, ER2 mg (Bydureon), copackaged8. Entacapone/Pramlintide: 1000 mg or 2000 mg tablet/600 mcg/mL (asacetate), copackaged9. Entacapone/Phentermine in 1000 mg/10 mg or 2000 mg/10 mg coformulatedtablets.10. Entacapone/atorvastatin (Lipitor) in 1000 mg/10 mg coformulatedtablets.11. Entacapone/fluvastatin (Lescol) in 1000 mg/80 mg coformulatedtablets.12. Entacapone/lovastatin (Altoprev, Mevacor) in 1000 mg/10 mgcoformulated tablets.13. Entacapone/pravastatin (Pravachol) in 1000 mg/10 mg coformulatedtablets.14. Entacapone/rosuvastatin (Crestor) in 1000 mg/5 mg coformulatedtablets.15. Entacapone/simvastatin (Zocor) in 1000 mg/10 mg coformulatedtablets.16. Entacapone/cholestyramine (Prevalite, Questran) in 1000/5000 mgcopackaged powder.17. Entacapone/colesevelam (Welchol) in 1000 mg/625 mg coformulatedtablets.18. Entacapone/colestipol (Colestid) in 1000 mg/1000 mg coformulatedtablets.19. Entacapone/ezetimibe (Zetia) in 1000 mg/10 mg coformulated tablets.20. Entacapone/ezetimibe-simvastatin (Vytorin) in 1000/10/10 mgcoformulated tablets.21. Entacapone/fenofibrate (Lofibra, TriCor) in 1000 mg/54 mgcoformulated tablets.22. Entacapone/gemfibrozil (Lopid) in 1000 mg/600 mg coformulatedtablets.23. Entacapone/Niacin (Niaspan) in 1000 mg/500 mg coformulated tablets.24. Entacapone/Omega-3 fatty acid (Lovaza) 1000 mg/4 mg coformulatedtablets.

By targeting different pathways the coadmininistered drugs can actsupplementally or synergistically to increase the potency compared withseparate administration, and thereby also permit use reduced orotherwise suboptimal or subtherapeutic dosages, if not coadministered,while maintaining efficacy, such as 50%/50% and 25%/25% coformulationsand copackagings of those shown in Table 2.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the invention containrelatively acidic functionalities, base addition salts can be obtainedby contacting the neutral form of such compounds with a sufficientamount of the desired base, either neat or in a suitable inert solvent.Examples of pharmaceutically acceptable base addition salts includesodium, potassium, calcium, ammonium, organic amino, or magnesium salt,or a similar salt. When compounds of the invention contain relativelybasic functionalities, acid addition salts can be obtained by contactingthe neutral form of such compounds with a sufficient amount of thedesired acid, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable acid addition salts include those derivedfrom inorganic acids like hydrochloric, hydrobromic, nitric, carbonic,monohydrogencarbonic, phosphoric, monohydrogenphosphoric,dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, orphosphorous acids and the like, as well as the salts derived fromrelatively nontoxic organic acids like acetic, propionic, isobutyric,oxalic, maleic, malonic, benzoic, succinic, suberic, fumaric, mandelic,phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric,methanesulfonic, and the like. Also included are salts of amino acidssuch as arginate and the like, and salts of organic acids likeglucuronic or galactunoric acids and the like. Certain specificcompounds of the invention contain both basic and acidic functionalitiesthat allow the compounds to be converted into either base or acidaddition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the invention.

In addition to salt forms, the compounds may be in a prodrug form.Prodrugs of the compounds are those compounds that undergo chemicalchanges under physiological conditions to provide the compounds of theinvention. Additionally, prodrugs can be converted to the compounds ofthe invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the invention when placed in a transdermal patch reservoirwith a suitable enzyme or chemical reagent. Prodrugs are often usefulbecause, in some situations, they may be easier to administer than theparent drug. They may, for instance, be more bioavailable by oraladministration than the parent drug. The prodrug may also have improvedsolubility in pharmacological compositions over the parent drug. A widevariety of prodrug derivatives are known in the art, such as those thatrely on hydrolytic cleavage or oxidative activation of the prodrug. Anexample, without limitation, of a prodrug would be a compound of theinvention which is administered as an ester (the “prodrug”), but then ismetabolically hydrolyzed to the carboxylic acid, the active entity.

Subject compounds can exist in unsolvated forms as well as solvatedforms, including hydrated forms. In general, the solvated forms areequivalent to unsolvated forms and are intended to be encompassed withinthe scope of the invention. Subject compounds may exist in multiplecrystalline or amorphous forms. In general, all physical forms areequivalent for the uses contemplated and are intended to be within thescope of the invention.

Certain subject compounds possess asymmetric carbon atoms (opticalcenters) or double bonds; the racemates, diastereomers, geometricisomers and individual isomers are all intended to be encompassed withinthe scope of the invention.

The compositions for administration can take the form of bulk liquidsolutions or suspensions, or bulk powders. More commonly, however, thecompositions are presented in unit dosage forms to facilitate accuratedosing. The term “unit dosage forms” refers to physically discrete unitssuitable as unitary dosages for human subjects and other mammals, eachunit containing a predetermined quantity of active material calculatedto produce the desired therapeutic effect, in association with asuitable pharmaceutical excipient. Typical unit dosage forms includeprefilled, premeasured ampules or syringes of the liquid compositions orpills, tablets, capsules, losenges or the like in the case of solidcompositions.

A wide variety of suitable formulations and delivery systems, includingsuitable excipients or carriers and methods for preparing administrablecompositions, are known or apparent to those skilled in the art and aredescribed in more detail in such publications as Remington: The Scienceand Practice of Pharmacy (Pharmaceutical Press (2012). For example, inparticular embodiments the compositions are formulated or delivered inextended or controlled delivery systems, such as diffusion systems (e.g.reservoir devices, matrix devices, diffusion-controlled implants andtransdermal patches) and encapsulated and matrix dissolution systems,erosion products, osmotic pump systems, ion exchange resins, etc.

In particular embodiment the amount administered is far in excess of, atleast 2, 2.5, 5 or 10× that (200 mg) currently indicated for Parkinson'sDisease, and will preferably be 0.5-10, 0.5-5, 0.5-2.5, 1-10, 1-5,1-2.5, 2-10, or 2-5 g/day, in unit dosage forms of 0.25, 0.5, 1, 1.5, 2or 2.5 g.

The compounds can be administered by a variety of methods including, butnot limited to, parenteral, topical, oral, or local administration, suchas by aerosol or transdermally, for prophylactic and/or therapeutictreatment. Also, in accordance with the knowledge of the skilledclinician, the therapeutic protocols (e.g., dosage amounts and times ofadministration) can be varied in view of the observed effects of theadministered therapeutic agents on the patient, and in view of theobserved responses of the disease to the administered therapeuticagents.

Examples Structure-Based Virtual Screening

We docked 1323 FDA-approved drugs to the substrate binding site in FTO,and filtered the docking poses based on physicochemical descriptors,such as the number of hydrogen bonds, buried carbon atoms andhydrophobic contact. Then, the docking poses were minimized and rescoredusing more sophisticated scoring method. One top-ranked drug isentacapone (FIG. 1), which is a COMT (Catechol-β-methyltransferase)inhibitor used for treating Parkinson disease.

Enzymatic Inhibition.

We measured the compound's inhibition activity in the demethylationreaction catalyzed by FTO. The assay was performed in 100 μl of reactionsystem containing 50 mM HEPES buffer (pH=7.0), 100 μM of a-KG, 100 μM of(NH₄)₂Fe(SO₄)₂, 1 mM of L-ascorbic acid, 50 ng/ml of BSA, 0.5 μM ofssDNA with N⁶-mA (5′-ATTGTCA(m⁶A)CAGCAGA-3′), and 0.1 μM of FTO protein.The reaction system was incubated at 37° C. for 2 h and stopped byheating at 95° C. for 5 mM ssDNA was digested by nuclease P1 andalkaline phosphatase. The concentrations of N⁶-mA and A were analyzed byHPLC-MS/MS. When concentration of substrate and enzyme are 0.5 μM and0.1 μM, respectively, the measured IC₅₀ value of entacapone against FTOis ˜9 μM (FIG. 2).

Cellular Triglyceride Synthesis Inhibition.

We measured the compound's inhibition activity on triglyceride synthesisin Huh-7 cell line (FIG. 3), the IC₅₀ value of entacapone is ˜15 uM.

In Vivo Anti-Obesity Efficacy.

Total of 23 male wistar rats (6 weeks) were fed with high-fat diet (45%fat, OpenSource Diets D12451), and entacapone (85.6 mg/day) wasadministered to 12 randomly selected rats by gavage. After 8 weeks, themean body weight of drug treatment group was about 11% less than that ofcontrol group (*p-value<0.05) (FIG. 4). However, thebody-weight-normalized food intakes of the two groups showed nodifference (FIG. 5). Interestingly, the LDL-c (Low DensityLipoprotein-cholesterol) in serum of drug treatment group decreased asabout 40% comparing to that of control group (***p-value<0.005) (FIG. 6and Table 1). The adipose and hepatic tissues of rat in drug-treatedgroup showed dramatic changes comparing to control group, with reducedsize of adipose cells and reduced level of liver steatosis (FIG. 7 a,bH&E staining, 10× magnification; c,d H&E staining, 40× magnification).

TABLE 1 Effects of Entacapone on serum biochemistry in rats. Differenceis the percentage of concentration change of index in drug treatmentgroup compared to that in control group. Index Difference p-valueCholesterol −3.3% 0.6455 HDL-cholesterol −0.026%  0.9972 LDL-cholesterol−41.2%  0.0005 Triglyceride −3.8% 0.8023 Bilirubin −10.1%  0.5814Albumin ratio −0.063%  0.9473 Glutamic-pyruvic transaminase −5.5% 0.4962Creatine kinase −17.6%  0.2311

From these and other data we determined that anti-weight gain effectivehuman dosages should be far in excess of the standard 200 mg/day dosagefor treating Parkinsons, preferably 0.5-10, 0.5-5, 0.5-2.5, 0.5-1, 1-10,1-5, 1-2.5, 2-10, or 2-5 g/day.

Atherosclerosis Model:

Ldlr-Deficient Mice. We measured entacapone's anti-atherosclerosisefficacy using Ldlr^(−/−) mice fed western style diet (20% fat, 0.15%cholesterol), entacapone (300 mg/day) was orally administered byblending with diet. After 8 weeks, the mean lesion area in aorticsinuses of drug treatment group was 28% less than that of control group(p-value=0.08) (FIG. 8). Therefore, entacapone reduces atherosclerosissymptom, comparing with the control.

In Vivo Anti-Obesity Efficacy in Obese Mice.

Male C57BL/6 mice were fed with high-fat diet (45% fat, OpenSource DietsD12451) for 8 weeks. Then obese mice with body weight 20% larger thanthat of mice fed with normal diet (20 mice) were selected forexperiments. Entacapone (300 mg/kg) was orally administered to 10randomly selected obese mice by blending with diet. After 13 weeks, themean body weight gain of drug treatment group was about 11% less thanthat of control group (*p-value<0.05) (FIG. 9).

REFERENCES

-   1. Frayling, T. M. et al. A common variant in the FTO gene is    associated with body mass index and predisposes to childhood and    adult obesity. Science 316, 889-894, (2007).-   2. Scuteri, A. et al. Genome-wide association scan shows genetic    variants in the FTO gene are associated with obesity-related traits.    PLoS Genet. 3, e115, doi:07-PLGE-RA-0253 (2007).-   3. Scott, L. J. et al. A genome-wide association study of type 2    diabetes in Finns detects multiple susceptibility variants. Science    316, 1341-1345, (2007).-   4. Dina, C. et al. Variation in FTO contributes to childhood obesity    and severe adult obesity. Nat Genet. 39, 724-726, (2007).-   5. Gao, X. et al. The fat mass and obesity associated gene FTO    functions in the brain to regulate postnatal growth in mice. PLoS    One 5, e14005, (2010).-   6. Fischer, J. et al. Inactivation of the Fto gene protects from    obesity. Nature 458, 894-898, (2009).-   7. Church, C. et al. A mouse model for the metabolic effects of the    human fat mass and obesity associated FTO gene. PLoS Genet. 5,    e1000599, (2009).-   8. Church, C. et al. Overexpression of Fto leads to increased food    intake and results in obesity. Nat Genet. 42, 1086-1092, (2010).-   9. Gerken, T. et al. The obesity-associated FTO gene encodes a    2-oxoglutarate-dependent nucleic acid demethylase. Science 318,    1469-1472 (2007).-   10. Han, Z. et al. Crystal structure of the FTO protein reveals    basis for its substrate specificity. Nature 464, 1205-1209, (2010).-   11. Meyer, K. D. et al. Comprehensive Analysis of mRNA Methylation    Reveals Enrichment in 3′ UTRs and near Stop Codons. Cell 149,    1635-1646 (2012).-   12. Jia, G. et al. N6-methyladenosine in nuclear RNA is a major    substrate of the obesity-associated FTO. Nat Chem Biol 7, 885-887,    (2011).

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein, including citations therein, are herebyincorporated by reference in their entirety for all purposes.

1-37. (canceled)
 38. A method of treating obesity or an obesity-relateddisease, comprising administering to a person in need thereof aneffective amount of entacapone((2E)-2-cyano-3-(3,4-dihydroxy-5-nitrophenyl)-N,N-diethylprop-2-enamide),or a pharmaceutically-acceptable salt thereof.
 39. The method of claim38 wherein the amount of entacapone or salt thereof is 0.5-5 g/day. 40.The method of claim 38 further comprising determining or detecting inthe person (a) genotype SNP rs7202116 (G), or (b) over-expression of FTOor Fto.
 41. The method of claim 38 further comprising detecting aresultant reduction in severity of the obesity or obesity-relateddisease.
 42. The method of claim 38 wherein the person is determined tobe in need of treatment to inhibit weight gain, promote weight loss,reduce serum LDL, cholesterol, LDL-c, or triglycerides, or treat anobesity related disease or ameliorate or reduce the pathology orseverity of an obesity related disease or a symptom of an obesityrelated disease, wherein the obesity related disease is selected fromdiabetes, hyperglycemia, diabetic nephropathy, hyperlipemia, coronaryheart disease, atherosclerosis, hypertension, cardiovascular orcerebrovascular diseases, and liver, kidney or thyroid diseases.
 43. Themethod of claim 38 wherein the person meets one or more criteriacontraindicative of Parkinson's disease, wherein the criteria are: (a)is not diagnosed with, or is not symptomatic of Parkinson's disease; and(b) is less than 50 years old.
 44. The method of claim 38 wherein theperson meets one or more criteria: (a) is obese or over-weight; (b)suffers from or is diagnosed with an obesity related disease selectedfrom diabetes, hyperglycemia, diabetic nephropathy, hyperlipemia,coronary heart disease, atherosclerosis, hypertension, cardiovascular orcerebrovascular diseases, or liver, kidney or thyroid diseases; (c) hasgenotype: SNP rs7202116 (G), rs1421085 (C), or rs9939609 (A), or asurrogate or proxy SNP in linkage disequilibrium therewith and having ar² value greater than 0.5; and (d) pathogenically expresses orover-expresses FTO or Fto.
 45. The method of claim 38 wherein: theamount of entacapone or salt thereof is 0.5-5 g/day; further comprisingdetermining or detecting in the person (a) genotype SNP rs7202116 (G),or (b) over-expression of FTO or Fto; further comprising detecting aresultant reduction in severity of the obesity or obesity-relateddisease; the person is determined to be in need of treatment to inhibitweight gain, promote weight loss, reduce serum LDL, cholesterol, LDL-c,or triglycerides, or treat an obesity related disease or ameliorate orreduce the pathology or severity of an obesity related disease or asymptom of an obesity related disease, wherein the obesity relateddisease is selected from diabetes, hyperglycemia, diabetic nephropathy,hyperlipemia, coronary heart disease, atherosclerosis, hypertension,cardiovascular or cerebrovascular diseases, and liver, kidney or thyroiddiseases; and the person meets one or more criteria contraindicative ofParkinson's disease wherein the criteria are: is not diagnosed with, oris not symptomatic of Parkinson's disease; and is less than 50 yearsold; and the person meets one or more criteria: is obese or over-weight;suffers from or is diagnosed with an obesity related disease selectedfrom diabetes, hyperglycemia, diabetic nephropathy, hyperlipemia,coronary heart disease, atherosclerosis, hypertension, cardiovascular orcerebrovascular diseases, or liver, kidney or thyroid diseases; hasgenotype: SNP rs7202116 (G), rs1421085 (C), or rs9939609 (A), or asurrogate or proxy SNP in linkage disequilibrium therewith and having ar² value greater than 0.5; and pathogenically expresses orover-expresses FTO or Fto.
 46. The method of claim 38 further comprisingadministering to the person an effective amount of a second, differentanti-obesity or anti-obesity-related disease medicament.
 47. The methodof claim 38 further comprising administering to the person an effectiveamount of a second, different anti-obesity or anti-obesity-relateddisease medicament, wherein: the amount of entacapone or salt thereof is0.5-5 g/day.
 48. The method of claim 38 further comprising administeringto the person an effective amount of a second, different anti-obesity oranti-obesity-related disease medicament: further comprising determiningor detecting in the person (a) genotype SNP rs7202116 (G), or (b)over-expression of FTO or Fto.
 49. The method of claim 38 furthercomprising administering to the person an effective amount of a second,different anti-obesity or anti-obesity-related disease medicament:further comprising detecting a resultant reduction in severity of theobesity or obesity-related disease.
 50. The method of claim 38 furthercomprising administering to the person an effective amount of a second,different anti-obesity or anti-obesity-related disease medicament,wherein: the person is determined to be in need of treatment to inhibitweight gain, promote weight loss, reduce serum LDL, cholesterol, LDL-c,or triglycerides, or treat an obesity related disease or ameliorate orreduce the pathology or severity of an obesity related disease or asymptom of an obesity related disease, wherein the obesity relateddisease is selected from diabetes, hyperglycemia, diabetic nephropathy,hyperlipemia, coronary heart disease, atherosclerosis, hypertension,cardiovascular or cerebrovascular diseases, and liver, kidney or thyroiddiseases.
 51. The method of claim 38 further comprising administering tothe person an effective amount of a second, different anti-obesity oranti-obesity-related disease medicament, wherein: the person meets oneor more criteria contraindicative of Parkinson's disease, wherein thecriteria are: (a) is not diagnosed with, or is not symptomatic ofParkinson's disease; and (b) is less than 50 years old.
 52. The methodof claim 38 further comprising administering to the person an effectiveamount of a second, different anti-obesity or anti-obesity-relateddisease medicament, wherein the person meets one or more criteria: (a)is obese or over-weight; (b) suffers from or is diagnosed with anobesity related disease selected from diabetes, hyperglycemia, diabeticnephropathy, hyperlipemia, coronary heart disease, atherosclerosis,hypertension, cardiovascular or cerebrovascular diseases, or liver,kidney or thyroid diseases; (c) has genotype: SNP rs7202116 (G),rs1421085 (C), or rs9939609 (A), or a surrogate or proxy SNP in linkagedisequilibrium therewith and having a r² value greater than 0.5; and (d)pathogenically expresses or over-expresses FTO or Fto.
 53. The method ofclaim 38 further comprising administering to the person an effectiveamount of a second, different anti-obesity or anti-obesity-relateddisease medicament, wherein: the second medicament is determined forinhibiting weight gain, promoting weight loss, reducing serum LDL,cholesterol, LDL-c, or triglycerides, or treating atherosclerosis. 54.The method of claim 38 further comprising administering to the person aneffective amount of a second, different anti-obesity oranti-obesity-related disease medicament, wherein: the second medicamentis determined for inhibiting weight gain, and is a food intake inhibitoror a food absorption inhibitor.
 55. The method of claim 38 furthercomprising administering to the person an effective amount of a second,different anti-obesity or anti-obesity-related disease medicament,wherein: the medicament is a determined for inhibiting weight gain, andis Orlistat, Sibutramine, Lorcaserin, Rimonabant, Metformin, Exenatide,Pramlintide, phentermine/topiramate, or a pharmaceutically-acceptablesalt thereof.
 56. The method of claim 38 further comprisingadministering to the person an effective amount of a second, differentanti-obesity or anti-obesity-related disease medicament, wherein: themedicament is determined for reducing serum LDL, cholesterol, LDL-c, ortriglycerides, and is atorvastatin (Lipitor), fluvastatin (Lescol),lovastatin (Altoprev, Mevacor), pravastatin (Pravachol), rosuvastatin(Crestor), simvastatin (Zocor), cholestyramine (Prevalite, Questran),colesevelam (Welchol), colestipol (Colestid), ezetimibe (Zetia),ezetimibe-simvastatin (Vytorin), fenofibrate (Lofibra, TriCor),gemfibrozil (Lopid), Niacin (Niaspan), Omega-3 fatty acid (Lovaza), or apharmaceutically-acceptable salt thereof.
 57. The method of claim 38further comprising administering to the person an effective amount of asecond, different anti-obesity or anti-obesity-related diseasemedicament, wherein: the entacapone or salt thereof, and secondmedicament are copackaged, coformulated or coadministered.